1.
Determination Of Comparative Effect Of Two Eeg Yolk Based Extenders On Post Thaw Semen Quality Of Sahiwal Bull
by Shahid Ali (2010-VA-05) | Prof. Dr. Main Abdul Sattar | Prof. Dr. manzoor Ahamd | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Cryopreservation of semen is the principal step for its usage in artificial insemination. Freezing of semen leads to reduction in post thaw semen quality. Glycerol has the cryoprotective properties that led to preserve spermatozoa. Egg yolk is also a basic constituent of semen extenders. Beneficial effect of egg yolk on sperm cryopreservation is plasma membrane protector. Tris based extenders are commonly used for semen cryopreservation of bulls, rams and bucks. Based on the economics and beneficial effects of extenders on bovine post thaw semen quality, tris based commercial as well as custom made semen extenders requiring egg yolk addition, needs to be considered for further studies. This study had been designed to determine the comparative effect of two egg yolk based extenders on post-thaw semen quality (Triladyl™and TCEY) on Sahiwal bulls. Semen was collected using an artificial vagina having temperature of 42 ºC from five adult Sahiwal regular donor bulls, raised at Center of Excellenc for Bovine Genetics (CEBG) Renalakhurd, Okara.Seven replicates of the experiment were performed. Volume, concentration and motility of ejaculates was evaluated. Semen samples having motility >60% and sperm/ml >500x106 were included in study. After evaluation, semen samples were pooled, divided into two aliquots of equal volume and kept in water bath at 37ºC. One aliquot was extended with tris citric acid egg yolk extender (TCEY) and other was extended with commercially available extender (Triladyl™). Pre-freeze CASA sperm motility parameters and kinematics of these extended aliquots were assessedat CEBG. After that, extended semen was cooled to 4 ºC for 4hr, equilibrated for 2hrs at 4ºC and packaged into 0.5 ml French straws (20 x 106 spermatozoa/straw). All semen straws were placed into automatic programmable freezer having liquid nitrogen vapors for 10 min. Afterward, shifted to liquid nitrogen for freezing and were stored until post-thaw semen evaluation carried out. The experiment was
repeated for seven times (replicates, n=seven). For post-thaw semen evaluation, four semen straws per treatment group were thawed (30 seconds) in water bath at 37ºC and post-thaw CASA sperm motility parameters and kinematics were checked. Post-thaw motility (PTM%) , Plasma Membrane Integrity (PMI %) , acrosome integrity (AI %) , live Sperm Percentage (LSP % )and sperm abnormalities (SA %) were checked by phase contrast microscope. Similarly AI (%), PMI (%), mitochondrial integrity (MI%) and DNA integrity (%) were checked by fluorescence microscope at Department of Theriogenology, University of Veterinary and Animal Sciences, Lahore.
Pre-freeze CASA sperm motility parameters;progressive %, rapid %and kinematics;average path velocity (VAP um/s), straight line velocity (VSLum/s), linearity (LIN %) were significantly better in Triladyl thanTCEY.Post thaw CASA sperm motility parameters; motile %, progressive %, rapid % and kinematics; VAP (um/s),VSL (um/s), straightness (STR %) and LIN (%) were also significantly better in Triladyl than TCEY. Post thaw semen quality parameters containing PTM (%), PMI (%), LSP (%), DI (%) andMI (%) were significantly better in Triladyl as compared to TCEY.
Availability: Items available for loan: UVAS Library [Call number: 2785-T] (1).
2.
Effect Of Adenosine 5’-Triphospate On Post Thaw Quality Of Buck Semen
by Ata-Ul-Rehman Hamaad (2015-VA-4340 | Prof. Dr. Main Abdul Sattar | Dr. Amjad Raiz | Dr.Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Livestock is the major source of income in agricultural countries. It boosts economic status by producing valuable food products and earning huge amount of Foreign exchange. Pakistan is rich in livestock population having more than 37 goat breeds. Artificial insemination is the most widely used ART and has made the most significant contribution to genetic improvement worldwide. It provides accurate means of progeny testing and spread of genetics to wide area with minimum cost. In contrast to its large scale use in bovine breeding, it is not routinely practiced in small ruminants. It may be due to effects of cryopreservation procedures on sperm plasma membrane, cytoskeleton, motility, mitochondrial membrane, acrosome and the nucleus of spermatozoa, which results in lower fertility rates and discourages the goat breeders to get benefit from this biotechnology. As mitochondria are power house of sperm cell, it provides energy to sperm cell and maintains motility. Therefore, the intactness of mitochondrial membrane must be maintained to keep the sperm cell motile after freezing and thawing procedure. In recent years, a pharmacologic agent, Adenosine 5’-triphosphate (ATP) has been used to enhance the quality and fertility of cryopreserved sperm. Previous reports have shown the freezing of mammalian sperm in hypertonic extenders to improve the rate of sperm viability and acrosome integrity. However, the impact of extracellular ATP supplementation on the quality of goat sperm after freezing and thawing still unknown. Thus the present study was conducted to evaluate the effects of Adenosine 5’ triphosphate (ATP) supplementation in pre-freeze and post thawed buck semen. The semen of six adult Beetal bucks was collected by artificial vagina maintained at 42ºC. After initial evaluation of percentage motility and concentration the semen was pooled and diluted with TCEY extender to the final concentration
Summary
42
of 200 × 106 sperms/mL. The study was conducted in two experiments, in first experiment the extended samples were divided in four equal aliquots and treated with one of the ATP concentrations (0 mM, 0.5 mM, 1 mM, 2 mM). All the samples were incubated at 35 ºC for 15 min. The samples were filled in 0.5 mL French straws cooled to 4ºC within 1 hr, equilibrated at 4oC for a period of 2hr and cryopreserved using standard freezing protocol. After freezing of minimum one week two straws from each aliquot were evaluated for percentages of post thaw motility, sperm viability, acrosome, plasma membrane, mitochondrial and DNA integrities. In second experiment the extended semen samples were cryopreserved with standard freezing protocols. Ten doses were thawed, evaluated individually for post thaw motility and pooled. Now the pooled samples was divided in four equal aliquots and treated with one of the ATP concentrations (0 mM, 0.5 mM, 1 mM, 2 mM). All the samples were incubated at 37 ºC and evaluated at 10 min, 1 hr, 2 hr, 3 hr and 6 hr for percentages of post thaw motility, sperm viability, acrosome, plasma membrane, mitochondrial and DNA integrities. The results of first experiment were analyzed by using one way ANOVA with SPSS. The results indicated that mitochondrial integrity % and DNA integrity % were higher (p < 0.05) in ATP treated groups. Sperm viability % was higher in 0.5 mM group as compared to all other groups. However, post thaw motility %, acrosome integrity % and plasma membrane integrity % remained similar in all groups. The results of second experiment were analyzed using GLM univariate procedure of SPSS. The results revealed that mitochondrial integrity % and DNA integrity % remained higher (p < 0.05) in ATP treated samples throughout the incubation time. Sperm viability % was higher (p < 0.05) in 2 mM ATP group while acrosome integrity % was higher (p < 0.05) in 1 mM ATP group throughout the incubation time. Post thaw motility % and plasma membrane integrity % remained similar (p > 0.05) throughout the incubation time. The interaction between time and
Summary
43
treatment was found non-significant (p>0.05). However, significant (p<0.05) decline in percentages of all parameters was observed with the increasing incubation time. In conclusion, addition of ATP enhances the post thaw quality of buck sperm by maintaining its motility, enhancing sperm viability, mitochondrial and DNA integrities. It also effects positively on post thaw quality of buck sperm. Moreover, is recommended in future to conduct research to elaborate the effects of ATP supplementation in semen on fertility of goats which is also primary objective of cryopreservation of semen. Availability: Items available for loan: UVAS Library [Call number: 2786-T] (1).
3.
Effect Of Resynchronization With Ovsynch Or Cidr On Cumulative Pregnancy And Embryonic Losses In Cidr-Gnrh Synchronized Nili-Ravi Buffalo
by Usman Arshad (2010-VA-235) | Prof. Dr. Nasim Ahmad | Prof. Dr. Main Abdul Sattar | Dr. Muhammad Hussan Saleem.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: The objective of the present study was to determine the effect of resynchronization with either GnRH or P4 on Day 23 (controlled internal drug release device containing progesterone; CIDR) on pregnancy rate, cumulative pregnancy, and embryonic and fetal losses in CIDR-GnRH synchronized Nili-Ravi buffaloes. Buffaloes (n = 181) of mixed parity, lactating, 181 ± 73 days postpartum, a body condition score (BCS) of 3.2 ± 0.5 (scale of 1-5), and 450-600 kg weight were subjected to synchronization and resynchronization. All buffaloes received CIDR on Day -9.5. In addition, GnRH was injected 36 h after CIDR removal, and fixed-time artificial insemination (FTAI) was performed 18 h later (Day 0). On Day 23, buffaloes were randomly assigned to receive one of the following treatments: 1) CON (n = 63), 2) P4 (n = 55), and 3) GnRH (n = 63) for resynchronization (2nd AI). Pregnancy rate and embryonic and fetal losses were monitored by serial ultrasonography on Days 30, 45, 60, and 90 after synchronization (1st AI), respectively. The pregnancy retention rate in GnRH-treated buffaloes remained significantly and consistently higher (P < 0.05) than in the CON group at Days 30, 45, 60, and 90 after 1st FTAI. Based on the pregnancy diagnosis, on Day 30 post 1st AI, buffaloes in the CON, P4, and GnRH groups received: 1) Artificial insemination on detected estrus (AIDE; n = 37), 2) CIDR-GnRH protocol (CIDR; n = 27), and 3) Ovsynch protocol (OVS; n = 23), respectively. The pregnancy rate in resynchronized buffaloes did not differ (P > 0.05) between the OVS and CIDR groups, whereas the cumulative pregnancy rate in GnRH + OVS buffaloes (81%) after the 1st and 2nd FTAI when determined on Day 64 was higher (P < 0.05) than that in CON + AIDE (59%) buffaloes. The embryonic losses were significantly lower (P < 0.05) in GnRH-treated (18%) buffaloes than in CON (42%) buffaloes on Day 45 post 1st AI. Fetal losses were fewer and did not differ (P > 0.05) due to treatments on Day 60 or 90 post- 1st AI. In conclusion, 1) the
Summary
28
pregnancy rate and cumulative pregnancy rate in GnRH + OVS buffaloes were higher than in CON + AIDE buffaloes when determined on Day 64 after synchronization and resynchronization and 2) embryonic and fetal losses were lower in GnRH-treated buffaloes than in CON buffaloes when determined from Day 31 - 90. Availability: Items available for loan: UVAS Library [Call number: 2787-T] (1).
